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. 2012 Aug 21;7(8):e43552. doi: 10.1371/journal.pone.0043552

Figure 6. ARL4D(T35N) overexpression does not cause cytochrome c release from mitochondria, induce apoptosis, or affect cell viability.

Figure 6

(A) At 36 h hours after transient transfection with plasmids encoding untagged ARL4D(T35N) or Bax-EGFP, COS-7 cells were fixed and incubated with antibodies against ARL4D or GFP plus cytochrome c. Bars, 10 µm. (B) GFP-Bax and the indicated ARL4D constructs were transiently expressed in COS cells. At 48 h after transfection, the cells were harvested, lysed and subjected to Western blot analysis with antibodies against ARL4D, GFP, PARP, or α-tubulin. *, cleaved PARP. (C) The viability of the RD cells was determined by MTT assay at 24, 48 and 96 h after transfection with plasmids encoding ARL4D or the indicated mutants. (D) RD cells were transfected with ARL4D, mutants, or an empty vector (EV). The stably transfected cells were subjected to G418 selection for two weeks, and the colonies remaining on the plates were stained with crystal violet (upper panel). The expression of ARL4D and its mutants after G418 selection was measured by Western blot (lower panel).